The Pigment Foundation of the Formation of Petal Stripes in the Rhododendron Variety Dayuanyangjin and the Exploration of Differential Expression Genes

doi:10.7668/hbnxb.20194067

Abstract

Abstract:

In order to analyze the molecular mechanism of the formation of pink stripes on the petals of Dayuanyangjin,a Rhododendron cultivar,and to explore the key genes related to the formation of pink stripes,the petals of Dayuanyangjin in its full bloom period were selected to determine the pigments in the white and pink stripe regions of the petals,and at the same time,their transcriptome was sequenced and analyzed using Illumina HiSeq^TM sequencing platform.Sequencing results were verified by Real-time Fluorescent Quantitative PCR (qRT-PCR).The results showed that the formation of pink stripes on the petals of Dayuanyangjin was caused by the accumulation of anthocyanins.A total of 7 086 differentially expressed genes (DEGs) were identified in the pink striped and white regions of petals,of which 3 802 were up-regulated and 3 284 were down-regulated.The anthocyanin biosynthesis,phenylpropanoid biosynthesis and flavonoid biosynthesis pathways related to anthocyanin formation were more active in the pink stripe region of petals.According to the results of the GO and KEGG enrichment analysis of DEGs,31 DEGs were screened from the related and regulatory pathways of anthocyanin biosynthesis,of which 26 were structural genes of anthocyanin biosynthesis pathway and 5 were transcriptional regulation genes (3 MYB and 2 bHLH);26 anthocyanin anabolic structure genes encode 9 enzymes,and the expression levels of 20 genes in the pink stripe tissue of petals were significantly higher than those in the white petal tissue.Through NCBI homology search,it was found that two R2R3-MYB and one bHLH were similar to the known homologous genes related to the regulation of anthocyanin synthesis in fruits or leaves,and their expression levels in the pink stripe region of petals were also significantly higher than those in the white region.Ten differentially expressed genes related to anthocyanin biosynthesis were selected and verified by qRT-PCR.

Key words: Rhododendron, Petal stripe, Anthocyanins, Transcriptome analysis, Differential expression genes, Transcription factor

SUN Xiaobo, GUO Zhenhao, LI Chang, ZHOU Huimin, LIU Xiaoqing, SU Jiale. The Pigment Foundation of the Formation of Petal Stripes in the Rhododendron Variety Dayuanyangjin and the Exploration of Differential Expression Genes[J]. Acta Agriculturae Boreali-Sinica, 2023, 38(6): 26-35. doi: 10.7668/hbnxb.20194067.

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Figures/Tables 11

References 41

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基因 Gene	基因号 ID	正向引物(5'—3') Forward primer(5'—3')	反向引物(5'—3') Reverse primer(5'—3')
Actin	KX230452	CGGTGCCCTGAAATCCTGTT	AAACGCTCAGCCATTCCAGG
MYB2	RHSIM_Rhsim08G0087200	GCACCAAGTCCCCCACAAA	CATCGTGCTCTGTGTCATTTCC
3GT	RHSIM_Rhsim04G0083600	AAGGAAACTAATGGAGAACGGG	GACAATCTTGCTCTTTTCGCTC
MYB31	RHSIM_Rhsim10G0036000	CGAGAATAATCTTTACGGTGGC	ATGGGTGAATCAGACGGGG
CHS	RHSIM_Rhsim08G0038400	GCTTGAAGGCACTTAGATGGAT	TGGCATTGTCTTATTGGTGATG
MYB17	DJ_newGene_2048	GTGCGGTAAGAGTTGTCGGC	TGTTGTCGTCTTCTGCGGG
FLS	RHSIM_RhsimUnG0095500	AGGATGACCGTTGGTATGATGT	CAAGGGAGAAAGTTCACTGAGG
DFR1	RHSIM_Rhsim08G0119200	TTTTACTGCTCCTACCCGCTAT	TTGAGGTCCGCCTTCCACA
DFR2	RHSIM_Rhsim06G0048400	TGTTGGAGGGTTGAGGAATGT	CTGTGACTTTTCCAGGACCGTA
F3H	RHSIM_Rhsim07G0247500	GCCTGTAAGTTGCTGGGGGT	TTGTGGGCATTTCGGGTAG
MYB15	RHSIM_Rhsim05G0060800	CGGGTTTCTTATTGGTGATGAC	TTCGGCAAAGGAATCCAACT

基因 Gene	基因号 ID	正向引物(5'—3') Forward primer(5'—3')	反向引物(5'—3') Reverse primer(5'—3')
Actin	KX230452	CGGTGCCCTGAAATCCTGTT	AAACGCTCAGCCATTCCAGG
MYB2	RHSIM_Rhsim08G0087200	GCACCAAGTCCCCCACAAA	CATCGTGCTCTGTGTCATTTCC
3GT	RHSIM_Rhsim04G0083600	AAGGAAACTAATGGAGAACGGG	GACAATCTTGCTCTTTTCGCTC
MYB31	RHSIM_Rhsim10G0036000	CGAGAATAATCTTTACGGTGGC	ATGGGTGAATCAGACGGGG
CHS	RHSIM_Rhsim08G0038400	GCTTGAAGGCACTTAGATGGAT	TGGCATTGTCTTATTGGTGATG
MYB17	DJ_newGene_2048	GTGCGGTAAGAGTTGTCGGC	TGTTGTCGTCTTCTGCGGG
FLS	RHSIM_RhsimUnG0095500	AGGATGACCGTTGGTATGATGT	CAAGGGAGAAAGTTCACTGAGG
DFR1	RHSIM_Rhsim08G0119200	TTTTACTGCTCCTACCCGCTAT	TTGAGGTCCGCCTTCCACA
DFR2	RHSIM_Rhsim06G0048400	TGTTGGAGGGTTGAGGAATGT	CTGTGACTTTTCCAGGACCGTA
F3H	RHSIM_Rhsim07G0247500	GCCTGTAAGTTGCTGGGGGT	TTGTGGGCATTTCGGGTAG
MYB15	RHSIM_Rhsim05G0060800	CGGGTTTCTTATTGGTGATGAC	TTCGGCAAAGGAATCCAACT

样品库 Library	过滤后的read数/条 Total Clean reads	过滤后碱基总数/Gb Total Clean bases	GC含量/% GC percentage	Q30/% Q30	比对基因组的read比例/% Ratio of reads mapped to the genome
W1	24 046 991	7.21	47.47	93.85	86.36
W2	22 503 598	6.75	47.09	93.37	85.48
W3	21 120 416	6.34	46.91	93.34	85.20
Y1	21 110 401	6.33	47.57	93.31	86.36
Y2	22 137 409	6.64	47.15	93.23	85.88
Y3	22 734 623	6.82	47.22	93.61	86.45

样品库 Library	过滤后的read数/条 Total Clean reads	过滤后碱基总数/Gb Total Clean bases	GC含量/% GC percentage	Q30/% Q30	比对基因组的read比例/% Ratio of reads mapped to the genome
W1	24 046 991	7.21	47.47	93.85	86.36
W2	22 503 598	6.75	47.09	93.37	85.48
W3	21 120 416	6.34	46.91	93.34	85.20
Y1	21 110 401	6.33	47.57	93.31	86.36
Y2	22 137 409	6.64	47.15	93.23	85.88
Y3	22 734 623	6.82	47.22	93.61	86.45

基因注释类别 Gene annotation category	代谢途径名称 Metabolic pathway name	富集因子 Enrichment factor	q值 q value	基因ID Gene ID
GO	生物过程:花青素生物合成过程的正调控(GO:0031542)	4.48	1	RHSIM_Rhsim08G0087600;RHSIM_Rhsim02G0231600; RHSIM_Rhsim07G0015600
GO	分子功能:花青素3-O-葡萄糖基转移酶活性(GO:0047213)	3.49	1	RHSIM_Rhsim06G0140700;RHSIM_Rhsim01G0008100; RHSIM_Rhsim06G0140800;RHSIM_Rhsim03G0263200
GO	生物过程:花青素生物合成过程的调节(GO:0031540)	1.84	1	RHSIM_Rhsim08G0087200;RHSIM_Rhsim08G0087300
GO	生物过程:含花青素的复合生物合成过程(GO:0009718)	1.79	1	RHSIM_Rhsim06G0048400;RHSIM_Rhsim08G0119200; RHSIM_Rhsim07G0096600
KEGG	花青素生物合成(map00942)	3.82	0.002 2	DJ_newGene_2880; RHSIM_Rhsim01G0007800; RHSIM_Rhsim01G0008100;RHSIM_Rhsim09G0150400; RHSIM_Rhsim09G0150500;RHSIM_Rhsim09G0150600; RHSIM_Rhsim03G0263200
KEGG	苯丙类生物合成(map00940)	1.76	1.95E-07	RHSIM_Rhsim01G0211500;RHSIM_Rhsim04G0145800; RHSIM_Rhsim12G0025100;RHSIM_Rhsim13G0096300; RHSIM_Rhsim09G0212900
KEGG	类黄酮生物合成(map00941)	1.61	0.018 0	RHSIM_Rhsim01G0013100;RHSIM_Rhsim06G0030400; RHSIM_RhsimUnG0066200;RHSIM_Rhsim07G0247500; RHSIM_Rhsim08G0033600;RHSIM_Rhsim08G0038400; RHSIM_Rhsim09G0036400;RHSIM_Rhsim09G0048100; RHSIM_Rhsim09G0048300

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